Synthetic and standardized prion infectuous material, and uses thereof as an injecting inoculum

Abstract

The invention concerns a novel, synthetic, standardised, soluble, reproducible and easy-to-handle infectious material of the prion type, consisting of a cell lysate or culture supernatant from stable transgenic cells expressing a prion protein PrP and supporting replication of the pathogenic form, PrPsc, of the said Prp.

1 . Cell lysate or culture supernatant obtained from stable transgenic cells expressing a prion protein PrP and supporting replication of the pathogenic form, PrPsc, of said PrP, whose infectious titer in PrPsc is greater than 50% of the infectious titer of a 10% homogenate of animals brains infected with said PrPsc. 
     
     
         2 . Cell lysate or culture supernatant obtained from stable transgenic cells expressing a prion protein PrP and supporting replication of the pathogenic form, PrPsc, of said PrP, whose infectious titer in PrPsc is greater than 75% of the infectious titer of a 10% homogenate of animals brains infected with said PrPsc. 
     
     
         3 . Cell lysate or culture supernatant obtained from stable transgenic cells expressing a prion protein PrP and supporting replication of the pathogenic form, PrPsc, of said PrP, whose infectious titer in PrPsc is greater than the infectious titer of a 10% homogenate of animals brains infected with said PrPsc. 
     
     
         4 . Cell lysate or culture supernatant according to  claim 1 , characterised by an infectious titer of ≧4 log TCID 50 /ml. 
     
     
         5 . Cell lysate or culture supernatant according to  claim 1 , characterised by an infectious titer of ≧5 log TCID 50 /ml. 
     
     
         6 . Cell lysate or culture supernatant according to  claim 1 , characterised by an infectious titer of ≧6 log TCID 50 /ml. 
     
     
         7 . Cell lysate or culture supernatant according to  claim 1 , characterised by an infectious titer of ≧7 log TCID 50 /ml. 
     
     
         8 . Cell lysate or culture supernatant according to  claim 1 , characterised in that the prion protein PrP expressed by the stable transgenic cells supporting the replication of the pathogenic form PrPsc is of ovine, bovine or human origin. 
     
     
         9 . Cell lysate or culture supernatant according to  claim 8 , characterised in that the prion protein PrP is the ovine prion protein of sheep Scrapie. 
     
     
         10 . Cell lysate or culture supernatant according to  claim 8 , characterised in that the prion protein PrP is the human prion protein of new-variant Creutzfeld-Jacob disease. 
     
     
         11 . Cell lysate or culture supernatant according to  claim 1 , characterised by the fact that the infected animals brains homogenate is obtained from the brains of animals of a given species infected with a PrPsc of the same species. 
     
     
         12 . Cell lysate or culture supernatant according to  claim 11 , characterised in that the infected animals brains homogenate is obtained from the brains of sheeps infected with sheep scrapie ovine PrPsc. 
     
     
         13 . Cell lysate or culture supernatant according to  claim 11 , characterised in that the infected animals brains homogenate is obtained from the brains of hamsters infected with hamster adapted PrPsc strain 263K. 
     
     
         14 . Cell lysate or culture supernatant according to  claim 11 , characterised in that the infected animals brains homogenate is obtained from the brains of cattle infected with bovine spongiform encephalitis (BSE) bovine PrPsc. 
     
     
         15 . Cell lysate or culture supernatant according to  claim 11 , characterised in that the infected animals brains homogenate is obtained from the brains of human patients infected with the human PrPsc of new-variant Creutzfeld-Jacob disease (vCJD). 
     
     
         16 . Cell lysate or culture supernatant according to  claim 1 , characterised in that the infected animals brains homogenate is obtained from the brains of transgenic animals expressing a PrP of a given species as a transgene and infected with a PrPsc of the same species. 
     
     
         17 . Cell lysate or culture supernatant according to  claim 16 , characterised in that the infected animals brains homogenate is obtained from the brains of transgenic mice expressing ovine PrP and infected with ovine PrPsc. 
     
     
         18 . Cell lysate or culture supernatant according to  claim 1 , obtained from stable transgenic epithelial rabbit cells. 
     
     
         19 . Cell lysate or culture supernatant according to  claim 18 , obtained from stable transgenic epithelial rabbit cell line Rov9. 
     
     
         20 . Cell lysate or culture supernatant according to  claim 1 , obtained from stable transgenic murine glial cells. 
     
     
         21 . Cell lysate or culture supernatant according to  claim 20 , obtained from stable transgenic murine glial cell line MovS6. 
     
     
         22 . Use of a lysate or culture supernatant according to  claim 1 , as an infecting inoculum for an evaluation and/or control method applied to a procedure used to obtain or process a biological product with the potential to be contaminated by a NCTA. 
     
     
         23 . Use of a lysate or culture supernatant according to  claim 22 , as an infecting inoculum for an evaluation and/or control method applied to a procedure used to purify blood plasma derivative products. 
     
     
         24 . Use of a lysate or culture supernatant according to  claim 23 , characterised in that the purification process implies chromatographies or nanofiltration. 
     
     
         25 . Use of a lysate or culture supernatant according to  claim 1 , as an infecting inoculum for an evaluation and/or control method applied to a procedure used to decontaminate equipment potentially contaminated with a NCTA. 
     
     
         26 . Use of a lysate or culture supernatant according to  claim 1 , as an infecting inoculum for a method evaluating a compound inhibiting the infectiousness of a NCTA. 
     
     
         27 . Use of a lysate or culture supernatant according to  claim 1 , as an infecting inoculum for an evaluation and/or control method for an infectious material confinement procedure.